Inhibitory effect of Cyrtomium falcatum on melanogenesis in α-MSH-stimulated B16F10 murine melanoma cells

Objective:To explore the anti-melanogenic potential of Cyrtomium falcatum.Methods:The effects of Cyrtomium falcatum crude extract and its solvent fractions on tyrosinase activity,melanin content,and the expressions of melanogenesis-related genes and proteins were analyzed inα-melanocyte-stimulating hormone(α-MSH)-stimulated B16F10 cells.Results:α-MSH treatment significantly increased tyrosinase activity,and extracellular and intracellular melanin content,as well as the expression levels of tyrosinase,microphthalmia-associated transcription factor(MITF),tyrosinase-related protein(TRP)-1,and TRP-2 in B16F10 cells.Treatment with Cyrtomium falcatum crude extract and its solvent fractions reduced tyrosinase activity and extracellular and intracellular melanin content and downregulated the expression levels of tyrosinase,MITF,TRP-1,and TRP-2 in a dose-dependent manner.Conclusions:Cyrtomium falcatum has potential anti-melanogenesis effects and can be used as a potential source material in cosmeceutical industry for the research and development of novel lead molecules with whitening properties.

Mushroom Tyrosinase Inhibition and Antimelanogenesis Activities of <i>Bacopa monnieri</i>(L.) Methanol Extract in B16F10 Melanoma Cells

Melanocytes that form stratum basale of skin epidermis express tyrosinase enzyme, which catalyzes initial two rate-limiting steps in the biotransformation of tyrosine into dark pigment called melanin. Even today, Tyrosinase inhibitors are among the promising candidates in cosmetic industry for skin-lightening formulations. Overexpression of tyrosinase causes excess melanin biosynthesis and deposition resulting in dark skin color. Moreover, localized overexpression of tyrosinase cause variety of hyperpigmentation disorders like melanoma, melasma, chloasma, dark patches, liver patches, etc. There has been a renewed interest in the natural products as main ingredients in the formulation of safe products for skin-whitening and treatment options for hyperpigmentation disorders. In the present communication, the results of our investigations on tyrosinase inhibition, modulation of intracellular tyrosinase and melanin levels in cultured B16F10 melanoma cells by Bacopa monnieri (L.) methanol extract (BME) are presented and discussed as safe option for skin lightening and to treat hyperpigmentation disorders. BME showed 11%, 29%, 54% and 80% inhibition of mushroom tyrosinase activity at an initial 100, 200, 400 and 600 μg of extract. Treatment of α-melanocyte stimulating hormone (α-MSH) stimulated cultured murine melanoma B16F10 cells with 100 μg/ml of the extract showed a decrease in the levels of cellular melanin and cellular tyrosinase content by 22% and 46% respectively. The cytotoxicity studies by MTT assay revealed that the LC50 of the BME is ≥1000 μg/ml in cultured mouse melanoma B16F10 and HEK293 cells.

Three-Dimensional Analysis of Melanosomes Isolated from B16 Melanoma Cells by Using Ultra High Voltage Electron Microscopy

Melanosomes, isolated by centrifugal separation from culture broth of B16 melanoma cells derived from mouse, were observed by scanning electron microscopy (SEM), and by transmission electron microscopy (TEM). Some interesting structural features were found inside and outside of the melanosomes. By SEM observation, the melanosomes were ellipsoid shape, their surface was not smooth and was covered with rough substructure, 10 to 20 nm particles. By TEM, uneven structure and micro particles were observed in the melanosomes. Furthermore, three-dimensional analysis was tried by using the ultra-high voltage electron microscopy(UHVEM). Micrographs of the melanosomes were taken at various tilted angles by UHVEM, after preparing 500 nm thickness specimens stained with lead citrate. From the micrographs collected, the three-dimensional structures were reconstructed by using i-mode software. Melanin stained by lead and non stained parts was clearly observed in the reconstructed structure. Non stained parts were round, regular size, and distributed widely in the melanosomes.

<i>In-Vitro</i>Effect of Sex Steroids on Mouse Melanoma (B16F10) Cell Growth

A battery of sex steroids were used to check their effect on mouse melanoma (B16F10) cell growth in-vitro. Progesterone and its synthetic receptor antagonist RU-486 showed maximum inhibition on in-vitro melanoma cell growth [1]. Further research work showed that the inhibition by progesterone was not a toxic, spurious or non-specific effect on mouse melanoma cell growth and the inhibition by progesterone was not mediated through progesterone receptor.

Influence of Ozonized Oil Nanoemulsions on B-16 Melanoma Cells: An <i>in Vitro Study</i>

In this study, we aimed to use a novel approach to overcome the current limitations of ozone therapy in medicine through ozonized oil nanoemulsions (OZNEs). We evaluated dose-dependency on the cellular activities of B-16 melanoma cell line which were incubated with various OZNE doses (v/v). Antitumor effects of OZNE against cancer cell lines were evaluated by cellular morphology, apoptosis and cell cycle analysis. Flow cytometry results showed that OZNE induced DNA damage, apoptosis, and arrested cell cycle in G0-1 phase in B-16 melanoma cells. Thus, OZNE treatment could pose an effective way to act as a potential therapeutic for patients with tumors in the future.

百合主要有效成分对小鼠B-16细胞黑色素含量、酪氨酸酶活性的影响

目的观察百合多糖与百合皂苷对小鼠B-16细胞中细胞活力、酪氨酸酶活性及黑色素含量的影响。方法选用同一传代小鼠B-16细胞并随机分为空白细胞组、空白对照组、熊果苷组、百合多糖组、百合皂苷组,共5组。空白细胞组加入180μL RPMI1640培养液;空白对照组加入180μL RPMI1640培养液和20μL高密度聚乙烯;熊果苷组、百合多糖组、百合皂苷组分别加入不同浓度(0.2、0.4、0.6、0.8 mg/mL)的熊果苷、百合多糖、百合皂苷,作用3 d后,用CCK-8法检测B-16细胞活力,用酪氨酸酶-多巴速率氧化法检测B-16细胞中酪氨酸酶的活性,以MTT法检测小鼠B-16细胞的黑色素含量。结果与空白细胞组比较,0.8 mg/mL熊果苷组细胞活力明显升高(P<0.01),0.4、0.6、0.8 mg/mL百合皂苷组细胞活力均明显降低(P<0.05,P<0.01);各浓度熊果苷、百合多糖组和0.6、0.8 mg/mL百合皂苷组酪氨酸酶活性均明显降低(P<0.01),0.2 mg/mL百合皂苷组酪氨酸酶活性明显升高(P<0.01)。与空白对照组比较,0.8 mg/mL熊果苷组细胞活力明显升高(P<0.01),0.6、0.8 mg/mL百合皂苷组细胞活力均明显降低(P<0.01);各浓度熊果苷、百合多糖组和0.6、0.8 mg/mL百合皂苷组酪氨酸酶活性均明显降低(P<0.01),0.2、0.4 mg/mL百合皂苷组酪氨酸酶活性均明显升高(P<0.01)。与0.2 mg/mL熊果苷组比较,0.8 mg/mL熊果苷组细胞活力、0.2 mg/mL百合皂苷组酪氨酸酶活性均明显升高(P<0.01)。与0.4 mg/mL熊果苷组比较,0.8 mg/mL熊果苷组细胞活力、0.4 mg/mL百合皂苷组酪氨酸酶活性均明显升高(P<0.01)。与0.6 mg/mL熊果苷组比较,0.6 mg/mL百合皂苷组酪氨酸酶活性均明显升高(P<0.01)。与0.4 mg/mL百合皂苷组比较,0.6、0.8 mg/mL百合皂苷组细胞活力均明显降低(P<0.05,P<0.01)。与0.6 mg/mL百合皂苷组比较,0.8 mg/mL百合皂苷组细胞活力明显降低(P<0.01)。在0.2、0.4 mg/mL浓度下,百合皂苷组酪氨酸酶活性均明显高于百合多糖组(P<0.01)。结论相较于百合皂苷、熊果苷,百合多糖能够在不影响小鼠B-16细胞活性的情况下,对B-16细胞内酪氨酸酶活性和黑色素含量起到抑制作用,推测百合多糖可能通过抑制细胞内酪氨酸酶的活性,进一步抑制黑色素的合成,达到美白的效果。

ENHANCED ANTITUMOR EFFECTS OF INTERLEUKIN-2 LIPOSOMES IN C57BL/6N MICE WITH B-16 MELANOMA PULMONARY METASTASES

为研究白细胞介素－２（ＩＬ－２）脂质体对小鼠Ｂ－１６黑色素瘤肺转移瘤的抑制效应和对荷瘤鼠脾淋巴细胞增殖的影响，应用改良超声－薄膜法制备ＩＬ－２脂质体；正常及免疫受抑Ｃ５７ＢＬ／６Ｎ小鼠建立Ｂ－１６黑色素瘤肺转移瘤模型，腹腔注射游离ＩＬ－２（０．２×１０４～５×１０４ｕｋｇ－１ｄ－１×１０ｄ）或脂质体包封的ＩＬ－２（０．０４×１０４～１．０×１０４ｕｋｇ－１ｄ－１×１０ｄ）。结果表明：改良超声－薄膜法制备的ＩＬ－２脂质体为直径２００～２５００ｎｍ的大单层脂质体，包封率４８．２％，具有良好的稳定性。ＩＬ－２脂质体可使荷瘤鼠肺湿重下降０．４％～１３．１％，肺转移瘤结节降低２．７％～４３．５％，脾细胞３Ｈ－ＴｄＲ掺入量增加１．０５～１．９０倍，其作用效应相当于５倍量的游离ＩＬ－２，且对免疫受抑荷瘤鼠的作用更强。提示ＩＬ－２经脂质体包封后，其抑瘤活性和促进淋巴细胞增殖的活性提高约５倍，是一种有希望应用于肿瘤生物治疗的新型制剂。

Liaoqiao aqueous extract inhibits B16 melanoma growth involving MAPKs/Nrf2/HO-1 mediated anti-oxidation and anti-inflammation

Aim Forsythia suspensa （Thunb.） Vahl, Lianqiao in Chinese, is one of the most fundamental herbs in traditional Chinese medicine （TCM） with heat-clearing and detoxicating properties. In this study, we aimed to study the antitumor activity of Lianqiao aqueous extract against melanoma using cancer cell line-based in vitro and mouse allografl tumor in vivo models. Furthermore, we also investigated the underlying molecular mechanisms, par- ticularly the involvement of anti-inflammation and anti-oxidation properties in its antitumor activity. Methods The proliferation of cancer cells was measured by MTT assay. The transplanted B16-F10 melanoma in C57BL/6 mice were established and used for the evaluation of in vivo antitumor effect of LQ. Tumor growth was monitored twice a week. Ki67 and CD31 were used to detect cancer cell proliferation and angiogenesis in tumor, respectively. The anti-oxidative property of LQ was determined by measuring the levels of ROS, MDA and GSH. The anti-inflamma- tory effect of LQ was evaluated by measuring TNF-α and IL-6 using ELISA kits. Other protein expression was deter- mined by Western Blot. Results LQ strongly inhibited the growth of B16-F10 cells in vitro and the tumor growth in vivo. The survival time of tumor-bearing mice was significantly prolonged by LQ. LQ inhibited cancer cell prolif- eration and angiogenesis in tumor as evidenced by decreased expressions of Ki67 and CD31. Levels of ROS, MDA TNF-α and IL-6 decreased, while GSH increased in LQ treatment group, indicating a strong anti-oxidative and an- ti-inflammatory activity of LQ. The expression of antioxidant proteins Nff-2 and HO-1, tumor suppressors P53 and p-PTEN, and the MAPK pathways in tumor tissues were upregulated by LQ treatment. Conclusions LQ exhibited strong antitumor activity against B16-F10 murine melanoma both in vitro and in vivo. The antitumor effect of LQ in- volved the decreased oxidative stress and inflammation in tumor, which is closely related to the heat-clearing and detoxicating properties of LQ.

程序性死亡配体1单克隆抗体增强DC疫苗致敏的B细胞靶向肿瘤干细胞的体液免疫

目的:探讨程序性死亡配体1(PD-L1)单克隆抗体是否可以增强ALDH high CSC-DC疫苗致敏的B细胞靶向ALDH high肿瘤干细胞(CSCs)的体液免疫作用。方法:建立B16-F10黑色素瘤小鼠模型,各组小鼠分别接受PBS、ALDH high CSC-DC+IgG、ALDH high CSC-DC疫苗、PD-L1单克隆抗体、ALDH high CSC-DC联合PD-L1单克隆抗体的治疗,记录小鼠的生存时间及肿瘤的体积。实验结束时收集各组小鼠的肿瘤,单个肿瘤细胞悬液进行ALDEFLUOR染色检测CSCs的比例。流式细胞术检测各组小鼠脾脏B细胞上PD-1的表达量。同时进一步行抗体结合试验和补体依赖的细胞毒性作用(CDC)试验检测B细胞培养上清中的抗体结合和裂解CSCs的能力。结果:相较于单独治疗组,PD-L1单克隆抗体与ALDH high CSC-DC疫苗的联合治疗可以更加显著地抑制肿瘤生长,延长小鼠生存时间。联合治疗组小鼠活化的B淋巴细胞上PD-1的表达水平显著降低,仅为6.5%。抗体结合试验提示,与ALDH high CSC-DC疫苗单独治疗组11.3%的结合率相比,联合治疗组小鼠B细胞培养上清中的抗体可以特异地结合15.7%ALDH high CSCs。同时CDC试验结果显示,联合治疗组的B细胞培养上清特异性地裂解ALDH high CSCs。结论:PD-L1单克隆抗体可以显著增强ALDH high CSC-DC疫苗致敏的B细胞产生靶向ALDH high CSCs的体液免疫反应。

Exposure of Weak Time-Invariant Electromagnetic Fields to B16-BL6 Cell Cultures Alter Biophoton Emission Profile as a Function of Distance

Biophoton emission is produced by all living systems;this emission pattern has been shown to be altered by the presence of an electromagnetic field (EMF). Cultures of B16-BL6 cells were exposed to a weak EMF produced by a specially constructed EM generator, called the “Resonator”, for one hour. This EM generator incorporates multiple geometric ratios in its design, including the golden ratio (phi), pi, root 2, root 3, and root 5. It has been used previously to purify water of toxins. There was a significant decrease in mean photon counts from B16-BL6 cells exposed at a distance of 1 m compared to those exposed at 0 m. Alterations in the spectral power density variability were also observed in the 8 - 10 Hz range. The EM generator may have an impact on the viability of the exposed cell cultures, but only at specific distances.

血清糖蛋白非转移性黑色素瘤蛋白B对表皮生长因子受体扩增伴随突变的非小细胞肺癌预后的预测作用

目的评估游离的糖蛋白非转移性黑色素瘤蛋白B(GPNMB)对表皮生长因子受体(EGFR)扩增伴随突变的非小细胞肺癌(NSCLC)患者的耐药和预后的预测价值。方法选取2018年3月至2019年9月在唐山市人民医院接受治疗的55例EGFR扩增伴随突变的NSCLC患者作为观察组,患者均应用EGFR酪氨酸激酶抑制剂(EGFR-TKI)作为一线治疗方案;随机选取同期体检中心67例健康人群血液样本作为对照组。比较2组游离GPNMB表达水平;采用t检验或χ2检验分析GPNMB表达和患者临床病理特征的相关性;结合临床疗效,评估其作为耐药标志物的价值。随访患者的无进展生存时间(PFS),并采用多因素Cox回归分析影响EGFR扩增伴随突变NSCLC患者生存的独立危险因素。结果与对照组相比,观察组游离GPNMB表达水平显著升高。观察组游离GPNMB水平和EGFR-TKI的临床疗效显著相关(P=0.016),GPNMB高表达患者的耐药程度更强,PFS也更短(P=0.032)。游离GPNMB高水平(HR=4.029,95%CI:1.942~8.358,P<0.001)是影响患者生存的独立危险因素。结论EGFR扩增伴随突变的NSCLC患者游离GPNMB表达水平显著上调;且其高表达与患者耐药程度的增强及不良预后显著相关,是影响患者生存的独立危险因素。

AURKB mRNA在皮肤黑素瘤中的表达及临床价值

目的探讨AURKB mRNA在黑素瘤组织中的表达及其与临床—病理特征及预后的关系。方法下载TCGA数据库中皮肤黑素瘤的数据集,采用COX回归分析皮肤黑素瘤患者不良预后的危险因素,AURKB mRNA表达水平与中位生存时间的关系采用GEPIA在线数据库分析,TIMER数据库分析皮肤黑素瘤组织中AURKB mRNA表达与免疫细胞浸润程度的相关性;皮肤黑素瘤中与AURB的共表达基因采用ULCAN数据库筛选,对共表达基因进行GO和KEGG富集分析。选择2例黑素瘤患者的肿瘤组织及癌旁组织,Western blot法验证AURKB在黑素瘤组织及癌旁组织中表达水平的差异。结果COX回归分析结果显示,病理学N分期(N2&N3)、Breslow深度(>3 mm)和AURKB mRNA(高表达)是皮肤黑素瘤患者发生不良预后的独立危险因素。皮肤黑素瘤组织中AURKB mRNA高表达228例,低表达229例,AURKB mRNA低表达组中位生存时间为102.82个月,高表达组中位生存时间为61.34个月,两组中位生存时间存在显著差异(HR=1.38,P=0.018)。在皮肤黑素瘤中,AURKB mRNA与Th2细胞(r=0.376,P<0.001)具有显著正相关性,与Th17细胞(r=-0.223,P<0.001)、NK细胞(r=-0.166,P<0.001)等具有显著负相关性。在皮肤黑素瘤中,与AURKB显著正相关和负相关的基因分别为646个和21个,选取100个显著正相关和21个显著负相关的基因进行富集分析,CC包括核浆、着丝粒、胞质等,MF包括蛋白质结合、微管结合等;BP包括细胞分裂等;KEGG包括细胞周期、DNA复制等。黑素瘤癌组织和癌旁组织中AURKB的相对灰度值分别为(1.332±0.214)和(0.321±0.021),与癌旁组织相比,AURKB表达水平在黑素瘤组织中显著升高(t=45.232,P<0.001)。结论AURKB在黑素瘤组织中表达水平显著升高,AURKB高表达与患者的不良预后有关,AURKB可能通过调控细胞周期等参与黑素瘤的恶性进展。

三氧化二砷诱导恶性黑色素瘤A_(375)和B_(16)细胞凋亡的研究

目的 :研究不同浓度的三氧化二砷 (As2 O3 )对恶性黑色素瘤人A3 75细胞和小鼠B16细胞的凋亡诱导作用 ,为As2 O3 治疗恶性黑色素瘤提供新的理论和实验依据。方法 :用流式细胞术 (FCM )检测A3 75细胞和B16细胞的凋亡诱导及杀伤作用 :用不同剂量的As2 O3 (0 .1mmol·L-1、0 .2 5mmol·L-1和 0 .5mmol/L-1) ,对小鼠进行腹腔注射 (10ml·kg-1,连续注射 10d后 ,检测小鼠血清与心、肝、肾有关的生化指标。结果 :As2 O3 浓度分别为 5 μmol·L-1、10 μmol·L-1和 2 0 μmol·L-1时 ,A3 75和B16细胞的凋亡率分别为 11.85 %、5 5 .5 1%、5 4 .90 %和 9.81%、2 6 .4 5 %、9.93% :As2 O3 诱导A3 75和B16细胞总的凋亡和坏死率分别为11.90 %、5 5 .71%、5 7.4 0 %和 12 .96 %、2 6 .99%、87.13% ;不同剂量的As2 O3 (0 .1mmol·L-1～ 0 .5mmol·L-1)对小鼠肝、肾和心肌等功能无明显影响 ,与对照组各项指标无显著性差异 (P >0 .0 5 )。结论 :不同浓度的As2 O3 能明显诱导恶性黑色素瘤A3 75及B16细胞凋亡和坏死 ,对A3 75细胞的凋亡诱导作用强于B16细胞。As2 O3 (0 .1mmol·L-1～ 0 .5mmol·L-1)对小鼠肝。

β-榄香烯对小鼠B_(16)黑色素瘤细胞粘附、运动和间隙连接通讯功能的影响

目的：探讨β揽香烯对小鼠Ｂ１６黑色素瘤细胞粘附和运动能力的影响。方法：采用细胞培养、电镜及划痕染料示踪技术，研究了β榄香烯对小鼠Ｂ１６黑色素瘤细胞粘附与运动能力、间隙连接结构及其间隙连接通讯功能的影响。结果：２０～４０μｇ／ｍｌ的β榄香烯可显著抑制Ｂ１６细胞对纤维结合蛋白和层粘连蛋白两种基质的粘附，抑制Ｂ１６细胞的运动，延迟吸管尖部所致细胞单层损伤的愈合。结论：提示β榄香烯对Ｂ１６细胞粘附和运动能力的抑制可能是其抗肿瘤转移作用的机制之一。

三氧化二砷对黑色素瘤B_(16)细胞端粒酶活性影响

目的 :研究三氧化二砷 (As2 O3 )对黑色素瘤B1 6细胞端粒酶活性的作用 ,探讨砷及其化合物治疗黑色素瘤的作用机制 ,为其治疗黑色素瘤提供新的理论和实验依据。方法 :用TRAP ELISA和TRAP PAGE银染法检测黑色素瘤B1 6细胞 (简称B1 6)端粒酶活性。结果 :As2 O3 对黑色素瘤B1 6细胞端粒酶活性的抑制有明显的浓度和时间依赖关系。结论 :As2 O3 对黑色素瘤B1

23-羟基桦木酸对B_(16)细胞系的诱导分化作用

目的评价 2 3 羟基桦木酸对黑色素瘤B16细胞的抑瘤作用。方法以MTT法测定细胞增殖的抑瘤率 ,并以B16细胞形态、黑色素含量、细胞周期变化及体内致瘤能力的测定作为观察指标。结果用 10～ 2 0 μg/ml的2 3 羟基桦木酸作用肿瘤细胞 ,见有不同程度的抑瘤作用 (P <0 .0 0 1)。表现为黑色素生成能力增加 ,细胞生长缓慢。可使B16细胞阻断在G1期 ,肿瘤体积明显缩小。结论 2 3 羟基桦木酸低剂量 (10～ 2 0 μg/ml)对B16细胞有明显的分化诱导作用 。

重组CCL2小鼠黑色素瘤B-16细胞的建立及生物学活性的鉴定

目的:在小鼠黑色素瘤B-16细胞中克隆表达趋化因子CCL2,以进一步研究趋化因子与肿瘤的关系。方法:从小鼠腹腔巨噬细胞中提取总RNA,RT-PCR扩增CCL2 cDNA,将此片段重组于真核表达载体pcDNA3.0,脂质体FuGENE6转染鼠黑色素瘤B-16细胞,G-418筛选阳性克隆。结果:RT-PCR和免疫细胞化学鉴定转染B-16细胞中有较强CCL2的表达,体外趋化实验表明重组CCL2有生物学活性。结论:获得有趋化活性的重组CCL2的小鼠黑色素瘤B-16细胞。

茯苓多糖对B16黑色素瘤人工肺转移模型的影响

[目的]探讨茯苓多糖对B16黑色素瘤小鼠人工肺转移的影响,并对其作用机制进行初步研究。[方法]C57BL/6小鼠接种B16黑色素瘤,分为茯苓多糖高、低剂量组、顺铂组、模型组,茯苓多糖高、低剂量组分别给予每只小鼠茯苓多糖0.5 mg、0.33 mg,顺铂组给予顺铂0.04 mg,模型组给予生理盐水,接种瘤细胞后第2天开始尾静脉注射给药,隔天给药1次。观察小鼠一般状态,造模后21 d取肺,计数肺表面转移灶个数、HE染色检测肺微小转移灶个数,并检测外周血白细胞数量及脾质量、脾指数。[结果]顺铂能够抑制肺转移,同时降低小鼠体质量、外周血白细胞数量、脾质量和脾指数。而茯苓多糖高、低剂量组对B16荷瘤鼠的体质量无明显影响,茯苓多糖高剂量可明显降低肺表面转移灶个数,茯苓多糖高、低剂量可减少肺微小转移灶个数,高剂量组外周血白细胞数量明显降低,低剂量组与模型组无统计学差异,低剂量可增加B16荷瘤鼠的脾质量和脾指数,高剂量对B16荷瘤鼠的脾质量和脾指数无明显影响。茯苓多糖高、低剂量组脾质量和脾指数均高于顺铂组。[结论]茯苓多糖能够抑制尾静脉注射B16荷瘤小鼠肺转移,增加其脾质量和脾指数,无明显增加其外周血白细胞数量的作用,推测活化外周血白细胞,增强免疫功能可能参与茯苓多糖抑制肿瘤转移的机制。

空间环境对黑色素瘤B16细胞致瘤基因和蛋白质表达的影响

初步探讨了空间环境对黑色素瘤B16细胞致瘤基因和蛋白质表达的影响。选择经第20颗返回式卫星搭载的B16细胞,进行体外和体内实验后,筛选出性状变异明显的空间诱变B16细胞株,检测空间诱变B16细胞基因和蛋白质表达的变化。将筛选出的空间诱变B16细胞接种于C57BL/6小鼠,2周后,处死荷瘤小鼠,取出移植瘤,利用免疫组化方法,检测接种空间诱变B16细胞的荷瘤小鼠移植瘤细胞Melan-A和VEGF蛋白的表达情况;利用RT-PCR方法,检测蛋白质水平上表达变化明显的空间诱变B16细胞melan-a,gp100,bag-3和l-pgds基因的表达情况。与对照细胞相比,1株空间诱变B16细胞荷瘤小鼠移植瘤细胞Melan-A和VEGF蛋白表达明显增加,其他细胞2种蛋白质的表达无明显变化。RT-PCR结果显示:此株细胞的melan-a,gp100和l-pgds基因表达增加,bag-3基因表达降低。空间环境的复合因素可诱导B16细胞基因和蛋白质表达产生变化。

海南文昌清澜港红树林真菌抗B16肿瘤细胞活性菌株的筛选

从海南文昌清澜港红树林采集78份植物组织和土壤样品。对样品进行真菌分离,共分离得到真菌608株,采用目测法对其体外抗肿瘤细胞活性进行检测,发现81株红树林真菌对小鼠黑色素瘤B16细胞有不同程度的抑制作用,占总分离菌株的13.32%。结果表明,从红树植物杯萼海桑中分离得到真菌菌株的数量最多,而从红树植物银叶树分离得到的抗肿瘤细胞活性菌株数量最多。